Agents and methods to elicit anti-tumor immune response
The invention provides an isolated, purified population of human cells comprising CD8 + T cells with reduced Cbl-b activity. The invention provides uses of such cells in methods for inducing or enhancing an anti-tumor immune response in a subject. These methods comprise: (a) providing a cell population, from a subject or from another source, which comprises CD8 + T cells, (b) reducing Cbl-b activity in the CD8 + T-cells, (c) administering the cells of step (b) to the subject. The invention provides methods for making CD8 + T cells that do not require stimulation through a co-receptor in order for the cell to become activated or proliferated in response to contact via its T cell receptor. Such methods are based upon reducing function of Cbl-b. The invention also provides methods for identifying agents which affect Cbl-b expression or activity.
1. An isolated, substantially purified, population of cells comprising CD8+ T cells with reduced Cbl-b activity compared to non-modified CD8+ T cells, wherein the Cbl-b activity is reduced by one or more of: (i) contacting the CD8+ T cells with a chemical agent which inhibits Cbl-b expression or activity, (ii) introducing an siRNA, which targets Cbl-b in the CD8+ T cells, or (iii) knocking out the gene encoding Cbl-b, and wherein the isolated, substantially purified, population of cells comprising CD8+ T cells with reduced Cbl-b activity compared to non-modified CD8+ T cells is included in a pharmaceutical composition.
2. The cells of claim 1 , wherein the CD8+ T-cells are provided from peripheral blood, lymph organs, or tumor infiltrates.
3. The cells of claim 1 , wherein the CD8+ T-cells are isolated from a subject.
4. The cells of claim 3 , wherein the CD8+ T-cells are isolated from tumor infiltrates from the subject.
5. The cells of claim 4 , wherein the CD8+ T-cells are polyclonal.
6. The cells of claim 1 , 2 , or 3 , wherein the CD8+ T cells with reduced Cbl-b activity have been stimulated to proliferate.
7. The cells of claim 1 , 2 , or 3 , wherein the CD8+ T cells with reduced Cbl-b activity have been stimulated to proliferate with tumor cells isolated from a subject, so as to increase the number of tumor specific CD8+ T cells with reduced Cbl-b activity.
8. The cells of claim 7 , wherein the tumor specific CD8+ T cells with reduced Cbl-b activity have been contacted with an anti-CD3 antibody or incubated with IL-2 or a combination thereof in order to further increase the number of the tumor specific CD8+ T cells with reduced Cbl-b activity.
9. The cells of claim 1 , 2 , or 3 , wherein the siRNA is SEQ ID NO: 1 (5′-CAGGAGTATGAGACAGAAG 3′).
10. The cells of claim 1 , wherein the pharmaceutical composition is used to treat cancer in a subject, or for immune rejection of a tumor in a subject.
11. The cells of claim 10 , wherein the subject suffers from melanoma, lymphoma, or any solid tumors expressing MHC-I with an antigen that can be recognized by cytotoxic T lymphocytes.